海洋科学2017，Vol.41Issue(9)：65-73,9.DOI:10.11759//hykx20170303001
牙鲆17β-HSD1基因克隆及其表达调控的初步研究
Molecular characterization, expression, and regulation of 17β-HSD1 in the olive flounder Paralichthys olivaceus
摘要
Abstract
17β-Hydroxysteroid dehydrogenase 1 (17β-HSD1) is an important enzyme as it is involved in the syn-thesis of both 17β-estradiol and testosterone. In this study,17β-HSD1 open reading frame sequence was obtained from the olive flounder Paralichthys olivaceus gonadal transcriptome data and verified. Phylogenetic tree analysis showed that flounder 17β-HSD1 was clustered with 17β-HSD1 proteins from fish species such as Oreochromis niloticus and Oryzias latipes. Semiquantitative RT-PCR results indicated the expression of 17β-HSD1 in the ovary was higher than that in the testis, and it was also expressed in the female gill, head kidney, kidney, spleen, stomach, and intestine. Real-time quantitative PCR analysis revealed upregulated expression patterns of17β-HSD1 during the key phases of ovarian and testicular differentiation.17β-HSD1 expression was significantly downregulated in the cultured primary testis cells treated with 75, 150, and 300μmol/L cAMP (P < 0.05). Moreover, transfecting the cul-tured primary testis cells with 2 and 3μg NR5a2 and 1μg NR0b1 significantly downregulated its expression, whereas it was significantly upregulated upon treatment with 2μg NR0b1 (P < 0.05), and all these regulations were dose-dependent. This study indicates that17β-HSD1 is involved in flounder gonadal differentiation, and its expres-sion is respectively regulated by cAMP, NR5a2, and NR0b1. These results may provide useful information for the study of fish gonadal differentiation.关键词
牙鲆(Paralichthys olivaceus)/17β-羟类固醇脱氢酶1/ORF克隆/表达/调控Key words
Paralichthys olivaceus/17β-HSD1/ORF clone/expression/regulation分类
生物科学引用本文复制引用
梁冬冬,范兆飞,邹玉霞,谭训刚,吴志昊,焦爽,李军,尤锋..牙鲆17β-HSD1基因克隆及其表达调控的初步研究[J].海洋科学,2017,41(9):65-73,9.基金项目
国家自然科学基金资助项目(41276171、31502156) （41276171、31502156）
青岛海洋科学与技术国家实验室鳌山科技创新计划项目(2015ASKJ02) （2015ASKJ02）
鲆鲽类产业技术体系课题(NYCYTX-50-G03) （NYCYTX-50-G03）
国家水产种质资源共享服务平台(2017DKA30470)National Natural Science Foundation of China, No. 41276171, 31502156 （2017DKA30470）
Qingdao National Laboratory for Marine Science and Technology, No. 2015ASKJ02 （）
The National Flatfish Industry System Construction Programme, No. NYCYTX-50-G03 （）
The National Infra-structure of Fishery Germplasm Resource , No.2017DKA30470 （）
